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Advances in sCMOS Camera Technology Benefit Bio Research - 6 Pages

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Catalogue excerpts

TECHNICAL NOTE Advances in sCMOS Camera Technology Benefit Bio Research sCMOS camera technology is gaining in popularity - Why? In recent years, cell biology has emphasized live cell dynamics, mechanisms and electrochemical signaling. As this research probes deeper into investigating rapidly changing phenomenon, the need for measuring high speed events at low light is constantly increasing. Scientific CMOS with its unique sensor architecture is able to provide both frame rates fast enough to capture high speed cell events and lower noise for better signal-to-noise measurements at the short exposure times required to achieve high frame rates. CCD and sCMOS comparisons, better or just different? Since the inception of digital microscopy, scientific grade CCD cameras have been the gold standard for imaging due to their sensitivity, linear response to light and low noise characteristics. However, many cell mechanisms occur on short time scales and emit low luminescence signals, making imaging with sufficient signal-to-noise and temporal and spatial resolution difficult for slower CCD cameras. Scientific CMOS is becoming the sensor technology of choice for these live cell fluorescence applications due to the ability of sCMOS to combine very low electronic noise with high frame rates and a large Field of View (FOV). As with anything, deciding whether a CCD or a sCMOS based camera is a better choice depends on the application. The prevailing factor to consider when deciding between a CCD or sCMOS camera is exposure time. Applications that require long exposure times ranging from a few minutes to a few hours are still best served by CCD cameras due to their lower dark current. These applications include bioluminescence and chemiluminescence from Western blot gels or in vivo animal imaging or electroluminescence from semiconducting materials. Fluorescence applications that can afford longer exposure times, including immunofluorescence of fixed cells, are well served by both sCMOS and CCD cameras. Considering these samples are not particularly sensitive to the effects of photobleaching, most researchers can afford to increase exposure times to several hundred milliseconds with an inexpensive CCD camera. That stated, the lower noise of sCMOS combined with its higher frame rates do provide higher quality images with shorter exposure times and a frame rate that makes it much easier to scan and focus on the sample. Fluorescence imaging of living cells on the other hand is extremely sensitive to light exposure. This is required to both minimize the photobleaching and phototoxicity effects as well as capture as much temporal information as possible. The objective is to capture images with sufficient signal-to-noise ratios while turning down the excitation intensity and using the shortest exposure time possible. sCMOS sensors offer a unique combination of low electronic noise that’s nearly one third of most high end interline CCD cameras with nearly 10x the frame rate potential. These two factors alone offer considerable advantages to traditional CCD cameras for most live cell fluorescence microscopy applications. ©2014 QImaging. All rights reserved.

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Advances in sCMOS Camera Technology Benefit Bio Research TECHNICAL NOTE How advances in sCMOS benefit bio research – What is optiMOS? optiMOS is a Scientific CMOS camera and CCD replacement, advanced to benefit bio research because it’s optimized specifically for fluorescence microscopy. Figure 1. Image and line profile comparison of the QImaging optiMOS sCMOS camera and a scientific grade cooled CCD camera. The image quality, signalto-noise levels, field of view and frame rates of the optiMOS are all superior to the scientific CCD camera. While CMOS sensors have been available for many years,...

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TECHNICAL NOTE Advances in sCMOS Camera Technology Benefit Bio Research Insufficient time resolution can result in two types of image artifacts: motion blur and temporal aliasing. Motion blur or streaking occurs when a selected exposure time is too long for a given velocity of a moving object. For example, imaging axonal transport of secretory granules involves tracking multiple particles as they move along a cell’s neurite with rapidly changing direction and velocity. If an exposure is long enough for a granule to either move a distance greater than a fraction of its diameter or change direction,...

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TECHNICAL NOTE Advances in sCMOS Camera Technology Benefit Bio Research In order to avoid any risk of aliasing, assume D is also equal to the length of the object. Given these three equations, it is possible to calculate the required frame rate for a given application to avoid motion blur and temporal aliasing. Below are a few examples: Object Size Cellular Event Object Size Object Velocity Required Frame Rates In-vivo red blood cell tracking 1ms exposures with >1000 frames per second (fps) to avoid motion blur and temporal aliasing Fast axonal transport of vesicles 3.3ms exposures with >60fps...

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Advances in sCMOS Camera Technology Benefit Bio Research TECHNICAL NOTE Lower cost of ownership, not compromised performance optiMOS is offered at a price associated with scientific interline CCD cameras but its low cost of ownership does not mean compromised performance. optiMOS offers cell biologists a high-speed, sensitive imaging solution that’s user-friendly, affordable, and bypasses the sometimes crippling data pains of rival sCMOS cameras. optiMOS does not however compromise on sensitivity nor frame rate as compared to alternative sCMOS options. Offering equivalent read noise levels of...

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Advances in sCMOS Camera Technology Benefit Bio Research TECHNICAL NOTE the supported optical field of view causing uneven illumination Moving field bottom right introduce spatial Unless a microscope is specifically designed to support large sensors, typically only the central region is used. Consequently, the benefit of the 5 megapixels cannot be exploited on many existing microscope configurations. Additionally, the 5 megapixel sCMOS cameras running at 10Ofps generate approximately 1.1 GB/s of data. In order to support this data stream, multiple SSD drives are required with a RAID 0 configuration...

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